The application of a sequential procedure with elimination as a method for the screening for metabolic diseases

A sequential classification procedure with early elimination, for the screening for metabolic diseases, is presented. Asymptotic properties of the procedure are derived in the Appendix and it is shown that the procedure is asymptotically distribution-free under certain assumptions, and asymptotically at least as efficient as a comparable fixed-sample procedure. With the use of data obtained from 36 mentally retarded patients, the procedure was evaluated by means of a bootstrap simulation. The procedure was then applied to this set of data, with satisfactory results and a considerable economy in observations.     

A multiple-tubes approach for accurate genotyping of very small DNA samples by using PCR: statistical considerations.

A multiple-tubes procedure is described for using PCR to determine the genotype of a very small DNA sample. The procedure involves dividing the sample among several tubes, then amplifying and typing the contents of each tube separately. The results are analyzed by a statistical procedure which determines whether a genotype can be conclusively assigned to the DNA sample. Simulation studies show that this procedure usually gives correct results even when the number of double-stranded fragments in the sample is as small as 30. The procedure remains effective even in the presence of small amounts of laboratory contamination. We find that the multiple-tubes procedure is superior to the standard one-tube procedure, either when the sample is small or when laboratory contamination is a potential problem; and we recommend its use in these situations. Because the procedure is statistical, it allows the degree of certainty in the result to be quantified and may be useful in other PCR applications as well.     

A two-step procedure for automatic and accurate segmentation of volumetric CLSM biofilm images

This paper presents a robust two-step segmentation procedure for the study of biofilm structure. Without user intervention, the procedure segments volumetric biofilm images generated by a confocal laser scanning microscopy (CLSM). This automated procedure implements an anisotropic diffusion filter as a preprocessing step and a 3D extension of the Otsu method for thresholding. Applying the anisotropic diffusion filter to even low-contrast CLSM images significantly improves the segmentation obtained with the 3D Otsu method. A comparison of the results for several CLSM data sets demonstrated that the accuracy of this procedure, unlike that of the objective threshold selection algorithm (OTS), is not affected by biofilm coverage levels and thus fills an important gap in developing a robust and objective segmenting procedure. The effectiveness of the present segmentation procedure is shown for CLSM images containing different bacterial strains. The image saturation handling capability of this procedure relaxes the constraints on user-selected gain and intensity settings of a CLSM. Therefore, this two-step procedure provides an automatic and accurate segmentation of biofilms that is independent of biofilm coverage levels and, in turn, lays a solid foundation for achieving objective analysis of biofilm structural parameters.     

Latissimus dorsi free flaps for total repair of extensive lower leg injuries in children

A well-established belief is that with crushed and contaminated wounds closure should be delayed. However, an emergency procedure involving very thorough debridement, complete reconstruction of all injured tissues, and cover by a latissimus dorsi free flap in the same operation is evaluated in 15 children presenting with severe injuries to the lower limb. It is felt that the procedure is superior to the established method because it is a one-stage procedure that minimizes the danger of infection, prevents growth impairment, shortens hospitalization, and allows early mobilization, thus being, in some cases, a limb-saving procedure.     

Stepwise Variable Selection in Classification Problems

Two variable selection procedures are evaluated for classification problems: a forward stepwise discrimination procedure, and a stepwise procedure preceded by a preliminary screening of variables on the basis of individual t statistics. Expected probability of correct classification is used as the measure of performance. A comparison is made of the procedures using samples from multi-variate normal populations and from several nonnormal populations. The study demonstrated some situations where the use of all variables is preferable to the use of a stepwise discriminant procedure stopping after a few steps, though usually the latter procedure was superior in performance. However where the stepwise procedure performed better than using all variables, the modified stepwise procedure performed still better. The use of modified stepwise procedures in which not all the covariances of the problem need be estimated seems promising.     

Regulatory affairs and biotechnology in Europe II.

This paper describes the EEC regulatory requirements for the preparation and execution of a community concertation "High Tech" procedure and compares this "High Tech" procedure with the Multi-State procedure. According to a decision of the European Commission enforced in July 1987, medicinal products, derived from high technology methods have been grouped in two categories: A. and B. Category A. concerns biotechnology products made by R-DNA techniques and by manipulation of mammalian cells. Category B. comprises all other products made by high technology. Before applying for an EEC marketing licence (e.g. submission for registration) one must ascertain whether a product is most appropriate in Category A. or B. and one should contact a licencing authority at an early stage to discuss the planned submission. Various procedures for submission have to be followed: 1. for the so-called "High Tech" products and especially products derived from biotechnology with therapeutic applications (Category A.), it is mandatory that one of the Member States accepts the submission. 2. The "High Tech" procedure is derived from the so-called "2-country" (Multi-State) procedure, in which for the latter procedure a marketing licence in one of the Member States (except Portugal) is required before application in other Member States. The Multi-State and "High Tech" (other products: Category B.) procedures are optional. When the procedures are started, all Member States concerned are involved in evaluation of full or abbreviated dossiers through mediation of the European Commission represented by the CPMP (Committee for Proprietary Medicinal Products), Brussels, Belgium. No application for a marketing licence of Category A. products is allowed without mediation of the CPMP. For Category B. products the applicant may opt for a national submission in one or more of the Member States without using the "High Tech" procedure. However, after consultation with the competent authority in one of the Member States, a "High Tech" procedure for Category B. products might still be advisable, but the applicant is not required to follow this procedure. Both the "High Tech" and the Multi-State procedure are currently executed by the mediation of a rapporteur, who liaises with the applicant from the start of the "High Tech" procedure. Ideally, the applicant should contact a licencing authority some 6 to 9 months before an application is planned: to ensure that the near future submission is acceptable. The institution of a rapporteur (appointed by the licencing authority in the country from where the procedure has recently been established) is introduced for the Multi-State procedure.(ABSTRACT TRUNCATED AT 400 WORDS)     

Graph selection with GGMselect

Applications on inference of biological networks have raised a strong interest in the problem of graph estimation in high-dimensional Gaussian graphical models. To handle this problem, we propose a two-stage procedure which first builds a family of candidate graphs from the data, and then selects one graph among this family according to a dedicated criterion. This estimation procedure is shown to be consistent in a high-dimensional setting, and its risk is controlled by a non-asymptotic oracle-like inequality. The procedure is tested on a real data set concerning gene expression data, and its performances are assessed on the basis of a large numerical study. The procedure is implemented in the R-package GGMselect available on the CRAN.     

Method for Measuring Mineralization in Lake Sediments

A method is described for measuring the mineralization of an organic solute ((14)C-glucose) by the heterotrophic indigenous bacteria in lake sediments. Since there is no suitable procedure for the determination of in situ microbial activities in sediments, the procedure described is probably the best devised so far and may serve as a base for a more definitive procedure.     

Detection of human papilloma virus DNA sequences by polymerase chain reaction

We have developed a polymerase chain reaction-based procedure for reproducible detection of the E6-E7 gene in human papilloma virus DNA sequences using formalin-fixed, paraffin-embedded tissue sections. This procedure is a simple one-step procedure which does not require any elaborate hybridization following polymerase chain reaction amplification. The protocol combines modified tissue treatment and proper primer selection for efficient amplification of target DNA in a highly specific manner allowing identification in ethidium bromide-stained gels. The procedure described here is useful for a variety of tissue preparations, particularly formalin-fixed, paraffin-embedded archival tissues.     

The determination of folylpolyglutamate synthetase.

A relatively rapid radiochemical procedure for the determination of folylpolyglutamate synthetase activity is presented in this communication. The procedure is based on measurement of the incorporation of radioactive l-glutamate into tetrahydropteroylpolyglutamate on incubation with a tetrahydrofolate. After deproteinating the incubation mixtures with trichloroacetic acid, folate is separated from radioactive glutamate by an adaptation of a procedure generally employed in the isolation of folate from natural materials, i.e., adsorption on columns of charcoal from which it is subsequently eluted with aqueous-alcoholic ammonia containing mercaptoethanol and counted. The procedure is applicable to monitoring purification of the enzymes and to the study of their properties.The technique for separating a radioactive product of enzyme action from a radioactive precursor with a column of charcoal, that has been developed for this procedure is applicable also to other radiochemical enzyme determinations requiring the separation of an aromatic from an aliphatic metabolite.     

生物大分子结晶学中的刚体修正方法

刚体修正方法是修正生物大分子初结构的取向和位置的一个很有效的方法.本文简要地介绍了Fourier搜索法,CORELS刚体修正法及TRAREF刚体修正法三种方法,并给出了它们在不同的蛋白质结构测定中应用的结果.     

Development of sequential multiple comparison procedure for dose response test

We propose a multiple comparison procedure to identify the minimum effective dose level by sequentially comparing each dose level with the zero dose level in the dose finding test. If we can find the minimum effective dose level at an early stage in the sequential test, it is possible to terminate the procedure in the dose finding test after a few group observations up to the dose level. Thus, the procedure is viable from an economical point of view when high costs are involved in obtaining the observations. In the procedure, we present an integral formula to determine the critical values for satisfying a predefined type I familywise error rate. Furthermore, we show how to determine the required sample size in order to guarantee the power of the test in the procedure. In practice, we compare the power of the test and the required sample size for various configurations of the population means in simulation studies and adopt our sequential procedure to the dose response test in a case study.     

Accurate and rapid estimation of phosphene thresholds (REPT)

To calibrate the intensity of transcranial magnetic stimulation (TMS) at the occipital pole, the phosphene threshold is used as a measure of cortical excitability. The phosphene threshold (PT) refers to the intensity of magnetic stimulation that induces illusory flashes of light (phosphenes) on a proportion of trials. The existing PT estimation procedures lack the accuracy and mathematical rigour of modern threshold estimation methods. We present an improved and automatic procedure for estimating the PT which is based on the well-established Ψ Bayesian adaptive staircase approach. To validate the new procedure, we compared it with another commonly used procedure for estimating the PT. We found that our procedure is more accurate, reliable, and rapid when compared with an existing PT measurement procedure. The new procedure is implemented in Matlab and works automatically with the Magstim Rapid(2) stimulator using a convenient graphical user interface. The Matlab program is freely available for download.     

Simultaneous One-Sided Pairwise Comparisons in a Two-Way Design

Cheung and Chan (1996) provided a procedure for simultaneous two-sided pairwise comparisons of treatment means in a two-way design. Expanding on this earlier work, we develop a procedure where all such comparisons are instead one-sided. The new procedure is appropriate in situations where the treatment means are a priori ordered. The overall type I error rate for the simultaneous inferences is controlled at a designated level. Tables of upper percentage points of the test statistic are given to facilitate the implementation of our method. The application of the testing procedure is illustrated with an example from a medical study.     

A Bayesian Decision Procedure for Selecting Prognostic Variables Associated with Survival for Data in which Censoring is Prevalent

A Bayesian procedure is developed for the selection of concomitant variables in survival models. The variables are selected in a step-up procedure according to the criterion of maximum expected likelihood, where the expectation is over the prior parameter space. Prior knowledge of the influence of these covariates on patient prognosis is incorporated into the analysis. The step-up procedure is stopped when the Bayes factor in favor of omitting the variable selected in a particular step exceeds a specified value. The resulting model with the selected variables is fitted using Bayes estimates of the coefficients. This technique is applied to Hodgkin's disease data from a large Cooperative Clinical Trial Group and the results are compared to the results from the classical likelihood selection procedure.     

A Selection and Testing Procedure for Comparing Several Experimental Treatments with a Control

We consider a selection and testing procedure for comparing k experimental treatments with a control treatment where the treatments are assumed to be normally distributed with unknown means and a common, unknown variance. Stein‐type sampling is used in the selection phase to screen for an experimental treatment that exhibits evidence of being better than the control treatment and each of the other experimental treatments, where better is defined in terms of the largest mean. In the testing phase, the best experimental treatment is compared to the control using a hypothesis test. If no experimental treatment indicates that it is an improvement over the control during the selection phase, our procedure allows for early termination. We provide definitions of level and power appropriate for our hybrid procedure and compute procedure parameters required to implement our procedure.     

Displacement imprinted polymer receptor analysis (DIPRA) for chlorophenolic contaminants in drinking water and packaging materials

The preparation of a molecularly imprinted polymer (MIP) for pentachlorophenol is described together with two alternative reporter derivatives for use in a displacement imprinted polymer receptor analysis (DIPRA) format procedure. In this procedure, alternative reporter molecules were rebound to the synthetic receptor sites and their displacement by the target analyte was employed as the basis of a simple procedure for the measurement of chlorophenols in water and packaging material samples. Water samples were extracted using the standard procedure (EPA 528) and a detection limit of 0.5 microg l(-1) was achieved using the DIPRA detection method, with good agreement between the displacement technique and GC-ECD analysis. A variety of packaging materials, extracted using a buffered detergent solution were also analysed using the DIPRA procedure and showed good agreement with GC results. In addition, investigation of the cross-reactivity of a range of pesticides and materials commonly encountered in environmental analysis indicated the procedure gave good discrimination between pesticides bearing a chlorophenolic moiety and other materials. The procedure is considered highly suitable for use as a rapid field-test method or for incorporation into a test kit device.     

Frequencies of Twelve Ascus-Types and Arrangement of Three Genes from Tetrad Data

Equations expressing the theoretical frequencies of twelve ascus-types in the tetrad analysis of a triply heterozygous diploid are described. Using these equations, a mapping procedure for a gene X, is proposed. The procedure requires that two genes, X and Y, of the same phenotype be heterozygous and that the map position of Y be known, and that another standard gene, Z, show an independent phenotype from X and Y. This procedure does not require the laborious allelism test of the segregants to determine the allelic 2:2 segregation in tetrads for the X and Y genes, which is indispensable for mapping by the conventional procedure. The exact placement of the X gene on a chromosome is possible by the chi2 minimization procedure in comparison with the expected frequencies of the six ascus-types or four spore-types deduced from the twelve expected ascus-types to give the optimal fit with the observed data.     

Quantum counter for correcting fluorescence excitation spectra at 320- to 800-nm wavelengths

A procedure for recording corrected fluorescence excitation spectra to wavelengths as long as 800 nm is described. The procedure involves the use of a commercial spectrofluorometer, which is modified by substituting 1,1',3,3,3',3'-hexamethylindotricarbocyanine perchlorate in place of rhodamine B as the quantum counter dye. This modification is applicable to spectrofluorometers supplied by several different manufacturers and can be accomplished by a user having only modest technical skills. A study of the fluorescence excitation spectrum of bacteriochlorophyll a is presented as an illustration of the use of the procedure. The procedure will be valuable in biological and biochemical studies that involve the use of long-wavelength fluorescent probes of either natural or synthetic origin.     

Stepwise Procedures under Unknown Variances for Toxicological Evaluation

Toxicological study is of practical importance in modern drug development. Proper statistical methodologies for toxicological evaluation of new developed drugs are undoubtedly necessary. In toxicological studies, it is practically desirable for a method to not declare the safety of a developed drug at a higher dosage prior to the declaration of the safety at lower dosages. Hsu and Berger 's stepwise confidence interval method was recently proposed for this purpose. Unfortunately, their procedure necessitates the homogeneity of variances among dosages, which is seldom satisfied in practice. In this article, via the application of the Stein 's two‐stage sampling method, we propose a stepwise confidence interval procedure for the same task without the homoscedasticity restriction. In addition, our procedure is shown to control its family‐wise type I error rate at the pre‐chosen nominal level. A simulation study will be conducted to compare our method, Hsu and Berger 's stepwise confidence interval method, and a single stage stepwise testing procedure based on Welch 's approximation. Our procedure is empirically shown to outperform Hsu and Berger 's procedure under heteroscedasticity and perform similarly with Welch 's procedure. An example will be used to illustrate our method.